Photolithographed micro-mirror well for 3d tomogram imaging of individual cells

Abstract

A micro-mirror well. In one embodiment the micro-mirror well includes a plurality of planar mirrors arranged around an axis of symmetry and inclined to form a pyramid well, where each of the plurality of planar mirrors is capable of reflecting light emitting from an object of interest placed inside the pyramid well.

What is claimed is: 
     
         1 . A process of fabricating a micro-mirror well, comprising the steps of:
 (a) providing a silicon substrate;   (b) etching off the silicon substrate to form an inverted pyramid well therein, wherein the inverted pyramid well has a plurality of side surfaces and a bottom surface extending to each other; and   (c) performing photolithographically masking and evaporating processes on the plurality of side surfaces and the bottom surface so as to form a mirrored pyramid well.   
     
     
         2 . The process of  claim 1 , wherein the etching step is performed with a potassium hydroxide (KOH) etching process. 
     
     
         3 . The process of  claim 1 , wherein each of the plurality of side surfaces is formed to define an angle, θ, relative to the bottom surface. 
     
     
         4 . A three-dimensional (3D) microscope, comprising:
 a numerical aperture (NA) objective having a plurality of mirrors configured to simultaneously collect images of an object of interest from multiple vantage points.   
     
     
         5 . The 3D microscope of  claim 4 , wherein the plurality of mirrors forms at least one mirrored pyramidal well (MPW). 
     
     
         6 . The 3D microscope of  claim 5 , wherein the at least one MPW has a plurality of side mirrored surfaces and a bottom mirrored surface, and wherein each of the plurality of side mirrored surfaces defines an angle, θ, relative to the bottom mirrored surface, and wherein 0°<θ<90°. 
     
     
         7 . The 3D microscope of  claim 5 , further comprising a microfluidic structure in communication with the at least one MPW. 
     
     
         8 . The 3D microscope of  claim 5 , wherein the at least one MPW is made from the smooth angled surfaces of anisotropically etched silicon. 
     
     
         9 . The 3D microscope of  claim 4 , wherein the object of interest comprises a biological analyte including cells and proteins. 
     
     
         10 . The 3D microscope of  claim 4 , wherein each of the plurality of mirrors comprises a dichroic mirror capable of reflecting specific wavelength ranges into a collection cone of the NA objective. 
     
     
         11 . The 3D microscope of  claim 4 , wherein the plurality of mirrors is affixed such that the perimeter of the field of view (FOV) of the NA objective contains reflected images of an object of interest. 
     
     
         12 . The 3D microscope of  claim 4 , wherein the plurality of mirrors is affixed opposite an object of interest from the NA objective for collecting reflected images of the object of interest. 
     
     
         13 . An algorithm for reconstruction of simultaneous, multi-vantage point images into three dimensional (3D) structures of an object of interest, comprising the steps of:
 (a) simultaneously collecting images of the object of interest form multi-vantage points surrounding the object of interest; and   (b) mapping the collected images of the object of interest to form a 3D image displaying the 3D structures of the object of interest.   
     
     
         14 . The algorithm of  claim 13 , wherein the step of simultaneously collecting images of the object of interest comprises the step of collecting light from simultaneously emitting fluorophores of the object of interest. 
     
     
         15 . The algorithm of  claim 14 , wherein the step of simultaneously collecting images of the object of interest is performed with a mirrored pyramidal well (MPW) having a plurality of side mirrored surfaces and a bottom mirrored surface, each of the plurality of side mirrored surfaces having an angle, θ, relative to the bottom mirrored surface, and wherein 0°<θ<90°.